Arsenic Trioxide and Acute Myeloid Leukemia

三氧化二砷与急性髓系白血病

• 批准号: 7770881
• 负责人: YONGKUI JING
• 金额: $ 25.51万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-15 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7770881
• 关键词: 4-ethoxymethylene-2-phenyl-2-oxazoline-5-one; Acids; Acute; Acute Myelocytic Leukemia; Acute Promyelocytic Leukemia; Aftercare; Animal Model; Apoptosis; Apoptotic; Arsenic Trioxide; Bacteria; Blood Cells; Blood specimen; Catabolism; Cell Death; Cell Line; Cells; Ceramides; Clinical; Clinical Trials; Combined Modality Therapy; Complex; Defect; Derivation procedure; Diuretics; Drug usage; Effectiveness; Enzymes; Esters; Ethacrynic Acid; Event; Fenretinide; Generations; Glutathione; Glutathione Reductase; Glutathione S-Transferase; Goals; HL60; Human; Hydrogen Peroxide; Hydroxyl Radical; In complete remission; Individual; Induction of Apoptosis; K562 Cells; Leukemic Cell; Luciferases; MCL1 protein; Malignant Neoplasms; Mediating; Membrane; Messenger RNA; Mitochondria; Mus; Myeloid Leukemia; NADPH Oxidase; NOD/SCID mouse; Organ; Pathway interactions; Patients; Peroxidases; Pharmaceutical Preparations; Physiological; Plasmids; Production; Proteins; Reactive Oxygen Species; Refractory; Relapse; Resistance; Respiration; Retinoids; Role; Sampling; Source; Sphingomyelins; Stratification; Superoxides; System; Tail; Testing; Therapeutic; Therapeutic Effect; Time; Toxic effect; Translating; Tretinoin; Tumor Burden; Weight; Xenograft procedure; analog; bioimaging; cancer cell; catalase; chemotherapy; clinical remission; design; efficacy testing; enzyme activity; glutathione peroxidase; human AKAP13 protein; in vivo; killings; leukemia; neutrophil; older patient; programs; public health relevance; response; retinamide; small hairpin RNA; success

DESCRIPTION (provided by applicant): The outstanding therapeutic success of As2O3 in a subtype of acute myelocytic leukemia (AML), the acute promyelocytic leukemia (APL), prompted widespread effort to extend this therapy to other malignancies. We and others determined that, through generation of reactive oxygen species (ROS), As2O3 induced apoptosis of APL cells. We also showed that degradation of PML-RARa was not a crucial event in the pro-apoptotic program of As2O3, but high ROS production ability was. We thus postulate that by enhancing the ROS-generating capacity through combination therapies we will be able to effectively target non-APL AML for apoptosis. Membrane localized NADPH oxidase and mitochondria are the main sources of ROS production. AML cells share lineage derivation with polymorphonuclear cells, which use NADPH oxidase generated ROS to kill bacteria. We showed that As2O3 treatment of AML cells increases the expression, without causing activation, of a crucial component of NADPH oxidase, p47phox. Our preliminary results show that a combination of therapeutically achievable concentration of As2O3 with fenretinide (4-HPR), a retinoid approved for clinical trials, synergizes in causing apoptosis in AML cells which express detectable (basal or As2O3-induced) NADPH oxidase components and extends survival of AML bearing SCID/NOD mice. Under physiological conditions ROS generated as a byproduct of mitochondrial respiration are detoxified by glutathione-enzymes, glutathione peroxidase (GPx), glutathione-s-transferase ? (GST?) and glutathione reductase (GR). Inhibition of the activities of these enzymes combined with increased ROS generation due to malfunction of the mitochondrial respiration in cancer cells, should generate levels of ROS that are beyond tolerable, causing mitochondria-mediated apoptosis of AML cells. Indeed, a combination of As2O3, which inhibits GPx, with ethacrynic acid (a clinically used diuretic drug) and its new derivative, which we have developed, and which inhibit GST? and GR, synergistically induce apoptosis in AML cells. This effect is independent of NADPH oxidase expression. We propose to study the mechanisms underlying these findings and their effect in primary AML and in vivo (animal models) in 4 specific aims. In aim 1 we will study the mechanism of As2O3 induced NADPH oxidase expression and its activation by 4-HPR through a ceramide dependent pathway. In aim 2 we will study the mechanism of the synergy of apoptosis induction by As2O3 and ethacrynic acid, and its derivative, through inhibition of ROS degradation and decrease of an antiapoptotic protein Mcl-1. In aim 3 we will study the apoptotic effects of As2O3 and 4-HPR or As2O3 and ethacrynic acid in primary AML samples. Importantly, we will investigate the possibility of using NADPH oxidase/myeloperoxidase or GST?/catalase levels in primary human AML cells as potential predictive markers of patient stratification to sensitive or resistant to As2O3/4-HPR or As2O3/ethacrynic acid therapies, respectively. In specific aim 4 we will test the efficacy of the combination treatments in xenografts of human AML. If further studies confirm that these treatment combinations are indeed effective in inducing apoptosis in AML cell lines and primary cells, and that they have therapeutic effect in xenografts, without excessive toxicity, because of the current clinical use of each of the tested compounds, it will be possible to rapidly translate these finding into clinical trials. PUBLIC HEALTH RELEVANCE: Arsenic trioxide (As2O3) is a drug used world-wide that induces complete clinical remission in 90% of APL patients without significant toxicity. Our therapeutic design includes the use of 4 HPR, a drug already in clinical trial, that activates a membrane system for the production of ROS and the use of new non-diuretic derivatives of ethyacrynic acid which block enzymes that diminish ROS and markedly enhanceAs2O3 induced leukemic cell death by several mechanisms. Our goal will be to utilize As2O3 in combination with 4 HPR or EA analogs in clinical trials in patients with refractory, secondary or in elderly patients with acute myelogenous leukemia.

描述（由申请人提供）：As 2 O3在急性髓细胞白血病（AML）的一种亚型（急性早幼粒细胞白血病（APL））中的显著治疗成功促使人们广泛努力将这种疗法扩展到其他恶性肿瘤。我们和其他人确定，通过产生活性氧（ROS），As 2 O3诱导APL细胞凋亡。我们还发现PML-RAR α的降解不是As_2O_3促凋亡程序中的关键事件，但高ROS产生能力是。因此，我们假设通过联合治疗增强ROS生成能力，我们将能够有效地靶向非APL AML细胞凋亡。膜定位的NADPH氧化酶和线粒体是ROS产生的主要来源。AML细胞与多形核细胞共享谱系衍生，其使用NADPH氧化酶产生的ROS来杀死细菌。我们发现，As 2 O3治疗AML细胞增加表达，而不引起激活，NADPH氧化酶，p47 phox的一个重要组成部分。我们的初步结果表明，治疗上可达到的浓度的As 2 O3与芬维A胺（4-HPR）（一种批准用于临床试验的类维生素A）的组合在引起表达可检测的（基础或As 2 O3诱导的）NADPH氧化酶组分的AML细胞凋亡方面具有协同作用，并延长了携带AML的SCID/NOD小鼠的生存期。在生理条件下，线粒体呼吸产生的活性氧被谷胱甘肽酶、谷胱甘肽过氧化物酶（GPx）、谷胱甘肽-S-转移酶（GST）、谷胱甘肽-S-转移酶（GST）、谷胱甘肽-S-转移酶（GST）和谷胱甘肽-S-转移酶（GST）解毒。(GST?)和谷胱甘肽还原酶（GR）。这些酶的活性的抑制与由于癌细胞中线粒体呼吸功能障碍而导致的ROS产生增加相结合，应该产生超出耐受范围的ROS水平，从而引起AML细胞的细胞凋亡。事实上，As 2 O3，抑制GPx，与依他尼酸（临床使用的利尿药）及其新衍生物，我们已经开发的组合，并抑制GST？和GR协同诱导AML细胞的凋亡。这种作用不依赖于NADPH氧化酶的表达。我们建议研究这些发现的机制及其在原发性AML和体内（动物模型）中的4个特定目标。目的一：研究As_2O_3诱导NADPH氧化酶表达的机制，以及4-HPR通过神经酰胺依赖途径激活NADPH氧化酶的机制。目的2：研究As_2O_3和依他尼酸及其衍生物通过抑制ROS降解和减少抗凋亡蛋白Mcl-1协同诱导细胞凋亡的机制。目的3：研究As_2O_3和4-HPR或As_2O_3和依他尼酸对原发性AML细胞凋亡的影响。重要的是，我们将研究使用NADPH氧化酶/髓过氧化物酶或GST？/原代人AML细胞中的过氧化氢酶水平分别作为患者分层为对As 2 O3/4-HPR或As 2 O3/依他尼酸疗法敏感或耐药的潜在预测标志物。在具体目标4中，我们将测试联合治疗在人AML异种移植物中的功效。如果进一步的研究证实这些治疗组合确实有效地诱导AML细胞系和原代细胞的凋亡，并且它们在异种移植物中具有治疗效果，而没有过度的毒性，则由于每种测试化合物的当前临床使用，将有可能将这些发现快速转化为临床试验。公共卫生关系：三氧化二砷（As 2 O3）是一种在世界范围内广泛使用的药物，可使90%的APL患者临床完全缓解，且无明显毒性。我们的治疗设计包括使用4 HPR，一种已经在临床试验中的药物，其激活用于产生ROS的膜系统，以及使用新的非利尿的乙炔酸衍生物，其阻断减少ROS的酶并通过几种机制显著增强As 2 O3诱导的白血病细胞死亡。我们的目标是在难治性、继发性或老年急性髓细胞性白血病患者的临床试验中利用As 2 O3与4种HPR或EA类似物联合。